Nitric oxide (NO) mediates cell signaling via cGMP- and non-cGMP-dependent reactions and yields secondary oxides of nitrogen (NOx) that expand the range of molecular targets of NO via oxidation, nitrosation and nitration reactions. The reactions of NO and its products with unsaturated fatty acids transduces NO signaling via the formation of electrophilic nitro-fatty acids, generically termed N02-FA. The research objectives set for the upcoming renewal of this R37/Merit award will continue to probe the concept that inflammatory-derived electrophilic fatty acids are redox signaling mediators that work in tandem with their highly conserved molecular targets (e.g., transcriptional regulatory proteins) to regulate adaptive cell signaling events. These reactions permit organisms to link gene expression with their metabolic, environmental and inflammatory status. Importantly, there still remains a lack of knowledge regarding the structural properties and biochemical reactivities of N02-FA that account for their induction of adaptive cell signaling responses. We will utilize HPLC-MS/MS to characterize the patterns and mechanisms of EFOX production during conditions relevant to obesity-induced diabetes. After learning more about the structural characteristics, concentrations, metabolism and molecular targets of specific EFOXs, we will evaluate the therapeutic potential of synthetic homologs of key EFOXs by administering these species in the setting of a model of obesity-induced diabetes, high fat diet-fed C57/BI6 mice. It is hypothesized that the electrophilic derivatives of unsaturated fatty acids formed by the oxidative inflammatory milieu mediate adaptive cell signaling responses. This hypothesis will be tested by pursuing three Specific Aims: 1. Structurally characterize and quantitate the predominant electrophilic fatty acid species present in the inflammatory milieu of obesity, using cell, rodent and clinically-derived specimens. 2. Define the contribution of mitochondrial redox reactions in the formation of electrophilic derivatives of unsaturated fatty acids. 3. Examine the anti-inflammatory actions of electrophilic fatty acids in cell and animal models of obesityinduced diabetes.

Public Health Relevance

The research plan capitalizes on the discovery that nitro- and keto-fatty acid derivatives are potent and broadly-acting anti-inflammatory mediators. We will a) identify new inflammatory-derived fatty acid electrophiles, and b) better understand the salutary cell signaling and physiological actions of these endogenously-produced electrophilic fatty acids and c) begin to develop these species as a new drug class.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37HL058115-17
Application #
8402190
Study Section
Special Emphasis Panel (NSS)
Program Officer
Charette, Marc F
Project Start
1998-01-15
Project End
2018-05-31
Budget Start
2013-07-25
Budget End
2014-05-31
Support Year
17
Fiscal Year
2013
Total Cost
$520,697
Indirect Cost
$177,062
Name
University of Pittsburgh
Department
Pharmacology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Ambrozova, Gabriela; Martiskova, Hana; Koudelka, Adolf et al. (2016) Nitro-oleic acid modulates classical and regulatory activation of macrophages and their involvement in pro-fibrotic responses. Free Radic Biol Med 90:252-60
Villacorta, Luis; Gao, Zhen; Schopfer, Francisco J et al. (2016) Nitro-fatty acids in cardiovascular regulation and diseases: characteristics and molecular mechanisms. Front Biosci (Landmark Ed) 21:873-89
Rudolph, Tanja K; Ravekes, Thorben; Klinke, Anna et al. (2016) Nitrated fatty acids suppress angiotensin II-mediated fibrotic remodelling and atrial fibrillation. Cardiovasc Res 109:174-84
Zhou, Chaoming; Ramaswamy, Swarna S; Johnson, Derrick E et al. (2016) Novel Roles for Peroxynitrite in Angiotensin II and CaMKII Signaling. Sci Rep 6:23416
Xie, Min; Mustovich, Anthony T; Jiang, Yi et al. (2015) IL-27 and type 2 immunity in asthmatic patients: association with severity, CXCL9, and signal transducer and activator of transcription signaling. J Allergy Clin Immunol 135:386-94
Wendell, Stacy Gelhaus; Golin-Bisello, Franca; Wenzel, Sally et al. (2015) 15-Hydroxyprostaglandin dehydrogenase generation of electrophilic lipid signaling mediators from hydroxy ω-3 fatty acids. J Biol Chem 290:5868-80
Snyder, Nathaniel W; Golin-Bisello, Franca; Gao, Yang et al. (2015) 15-Oxoeicosatetraenoic acid is a 15-hydroxyprostaglandin dehydrogenase-derived electrophilic mediator of inflammatory signaling pathways. Chem Biol Interact 234:144-53
Fazzari, Marco; Khoo, Nicholas; Woodcock, Steven R et al. (2015) Generation and esterification of electrophilic fatty acid nitroalkenes in triacylglycerides. Free Radic Biol Med 87:113-24
Vitturi, Dario A; Minarrieta, Lucia; Salvatore, Sonia R et al. (2015) Convergence of biological nitration and nitrosation via symmetrical nitrous anhydride. Nat Chem Biol 11:504-10
Delmastro-Greenwood, Meghan; Hughan, Kara S; Vitturi, Dario A et al. (2015) Nitrite and nitrate-dependent generation of anti-inflammatory fatty acid nitroalkenes. Free Radic Biol Med 89:333-41

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